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Article|01 Aug 2021|OPEN
Illuminating the cells: transient transformation of citrus to study gene functions and organelle activities related to fruit quality
Jinli Gong1,2,3, Zhen Tian1,2,3, Xiaolu Qu1, Qiunan Meng1,2,3, Yajie Guan1,2,3, Ping Liu4, Chuanwu Chen4, Xiuxin Deng1, Wenwu Guo1, Yunjiang Cheng1,2 & Pengwei Wang2,3,1,
1Key Laboratory of Horticultural Plant Biology (Ministry of Education), College of Horticulture and Forestry Science, Huazhong Agricultural University, 430070 Wuhan, China
2National R&D Centre for Citrus Preservation, Huazhong Agricultural University, 430070 Wuhan, China
3 Interdisciplinary Sciences Research Institute, Huazhong Agricultural University, 430070 Wuhan, China
4Guangxi Academy of Specialty Crops/Guangxi Engineering Research Center of Citrus Breeding and Culture, 541004 Guilin, China

Horticulture Research 8,
Article number: 175 (2021)
doi: 10.1038/hortres.2021.175
Views: 121

Received: 11 Jan 2021
Revised: 14 Apr 2021
Accepted: 04 May 2021
Published online: 01 Aug 2021

Abstract

Although multiple microscopic techniques have been applied to horticultural research, few studies of individual organelles in living fruit cells have been reported to date. In this paper, we established an efficient system for the transient transformation of citrus fruits using an Agrobacterium-mediated method. Kumquat (Fortunella crassifolia Swingle) was used; it exhibits higher transformation efficiency than all citrus fruits that have been tested and a prolonged-expression window. Fruits were transformed with fluorescent reporters, and confocal microscopy and live-cell imaging were used to study their localization and dynamics. Moreover, various pH sensors targeting different subcellular compartments were expressed, and the local pH environments in cells from different plant tissues were compared. The results indicated that vacuoles are most likely the main organelles that contribute to the low pH of citrus fruits. In summary, our method is effective for studying various membrane trafficking events, protein localization, and cell physiology in fruit and can provide new insight into fruit biology research.